Age-related macular degeneration (AMD) is a complex disease and the leading cause of blindness in the elderly. Many individuals carrying AMD risk genotypes never develop the disease and only a fraction of those diagnosed with AMD progress to advanced disease with severe visual loss. Ophthalmologists cannot determine which patients will progress to advanced AMD. Our long-term goal is the development of prognostic technology for assessing advanced AMD risk and for monitoring AMD therapeutics. Our previous biomarker analyses have confirmed the AMD biomarker potential of plasma protein oxidative modifications such as carboxyethylpyrrole (CEP), carboxymethyllysine (CML) and pentosidine and that proteomic and genomic AMD biomarkers are more effective when used together than alone. We propose two exploratory aims that test the hypothesis that CEP-derivatives of ethanolamine phospholipids (CEP-EPs) are superior AMD biomarkers compared to CEP-protein modifications. Our preliminary studies suggest that like CEP-protein, CEP-EPs induce angiogenesis through toll-like receptor 2. Furthermore we have developed methodology for the efficient conversion of a multitude of CEP-EPs to a single derivative, namely CEP-ethanolamine (CEP-ETN) and have established a LC MS/MS method for quantifying CEP-ETN in plasma. Our pilot study measuring plasma CEP-ETN indicates CEP-EPs are much more elevated in AMD than normal plasma, more so than CEP-protein. In aim 1, we will validate by LC MS/MS elevated CEP-ETN in AMD plasma and demonstrate AMD biomarker utility both alone and in combination with genomic AMD markers and CEP-protein, CML and pentosidine. The results will enhance the discriminatory accuracy of AMD biomarkers. In aim 2, we will confirm that CEP-EP derivatives are elevated in AMD ocular tissues using MALDI imaging mass spectrometry, and establish quantifiable endpoints for future animal studies evaluating AMD therapeutics. Outstanding resources include a large, well-characterized plasma repository, the experience of the investigative team and a world-class vision research environment. The overall impact will be high, as effective biomarkers will transform AMD patient management.